Effect of electrochemotherapy with betulinic acid or cisplatin on regulation of heat shock proteins in metastatic human carcinoma cells in vitro

Mączyńska,Justyna; Choromańska,Anna; Kutkowska,Justyna; Kotulska,Małgorzata; Zalewski,Maciej; Zalewski,Jerzy; Kulbacka,Julita; Saczko,Jolanta

doi:10.3892/or.2019.7103

Effect of electrochemotherapy with betulinic acid or cisplatin on regulation of heat shock proteins in metastatic human carcinoma cells in vitro

Authors:
- Justyna Mączyńska
- Anna Choromańska
- Justyna Kutkowska
- Małgorzata Kotulska
- Maciej Zalewski
- Jerzy Zalewski
- Julita Kulbacka
- Jolanta Saczko
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Affiliations: Division of Radiotherapy and Imaging, Institute of Cancer Research, Sutton SM2 5NG, UK, Department of Medical Biochemistry, Wroclaw Medical University, 50‑368 Wroclaw, Poland, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 53‑114 Wroclaw, Poland, Institute of Biomedical Engineering and Instrumentation, Wroclaw University of Science and Technology, 50‑370 Wroclaw, Poland, Department of Gynecology and Obstetrics, Wroclaw Medical University, 51‑618 Wroclaw, Poland, Department of Molecular and Cellular Biology, Faculty of Pharmacy with Division of Laboratory Diagnostics, Wroclaw Medical University, 50‑556 Wroclaw, Poland
Published online on: April 8, 2019 https://doi.org/10.3892/or.2019.7103
Pages: 3444-3454

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Abstract

Betulinic acid (BTA) is naturally occurring triterpene that has received interest as a novel therapeutic substance with cytotoxicity towards a number of cancer cell lines. Despite the wide spectrum of biological and pharmacological effects, its effect may be limited its lipophobic properties. Therefore, strategies to improve the access of BTA to the cells are required to enhance the anticancer effects. Electroporation (EP) enables increased inflow of drugs into cancer cells, even at low doses, which may reduce the side effects caused by high doses of chemotherapy. The potential application of BTA in electrochemotherapy (ECT) in metastatic type of cancers was investigated in the present study. The efficacy of BTA with EP was estimated using a cell survival assay (MTT assay), microscopical morphology analysis and the immunocytochemical expression of heat shock proteins (HSPs). HSPs are molecules that protect the cell from harmful environmental, chemical and physical stresses, and ensure cell survival, recovery and proper functioning. HSP expression is induced various stress factors. Therefore, the expression of HSP27 and HSP70 was evaluated after cells were exposed to an external pulsed electric field and anticancer drugs. Facilitated drug delivery and the anticancer effect on metastatic tumor cells were evaluated in vitro. The effect of BTA was compared with cisplatin (CP), a standard cytostatic agent. Two different metastatic cancer cell lines were used, an ovary adenocarcinoma cell line (SW626) and melanoma cell line (Me45). BTA combined with EP exhibited similar efficacy to CP with EP after 24 and 48 h in SW626 and Me45 cancer cells. Me45 cells also had high HSP27 and low HSP70 immunosignals post‑ECT treatment. ECT caused increased expression of HSP27 and HSP70 proteins in SW626 cells, which were less sensitive to ECT than the Me45 melanoma cell line. The results indicate that BTA may be efficiently applied instead of CP in ECT approaches, but its activity differs between tumor cell lines.

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